Subcellular proteomics reveals a blueprint for endosymbiont integration in trypanosomatid Angomonas deanei.

The acquisition of endosymbionts is a fundamental process that has driven the evolution of eukaryotes. The tree of life is filled with cases of internalised prokaryotes that have become integrated into their hosts, often forming mutually beneficial relationships. The trypanosomatid Angomonas deanei is one such case, harbouring a single β-proteobacterial endosymbiont. This symbiotic relationship is highly advanced, as evidenced by the identification of host-encoded proteins that are targeted to the bacterium and control its division. To deeper understand this integration, we performed an in-depth subcellular proteomic analysis to determine the compartmental localisation of both host and endosymbiont proteins. Our analysis resolved over 5,000 host proteins and over 400 endosymbiont proteins. We used this rich dataset to identify several novel host-encoded proteins targeted to the bacterium, and validated our predictions using genetic manipulations and microscopy. By mapping the localised enzymatic repertoire, we were able to shed light on metabolic interplay between the two organisms. We confirmed an energetic basis for the previously observed association between the host's glycosomes and its endosymbiont, and discovered an interaction between the endosymbiont and the host's acidocalcisomes. This subcellular proteomic dataset provides a comprehensive foundation for future research into the remarkable process of bacterial integration.